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Project grant

The human tumour micro-environment modelled in in vitro biomatrices and applied to cancer drug discovery

a photo showing a glass slide being held by a scientist, near to a microscope

At a glance

Completed
Award date
October 2009 - February 2013
Grant amount
£407,235
Principal investigator
Dr Anna Grabowska

Co-investigator(s)

Institute
University of Nottingham

R

  • Replacement
Read the abstract
View the grant profile on GtR

Application abstract

The paracrine signalling pathways within a tumour micro-environment play an important role in the epithelial–mesenchymal transition (EMT) and metastasis. Cancer-associated fibroblasts (CAFs), in particular, have important roles in supporting these processes. Stromal–epithelial interactions have become a focus of cancer drug discovery but are not optimally modelled by xenograft systems. For example, transplanted human epithelial cells are infiltrated with murine stroma but within the hepatocyte growth factor (HGF)/C-met axis, an important paracrine signalling pathway, murine HGF from the stroma does not bind the human C-met receptor on human epithelial cells within xenografts. Therefore to replace the inappropriate use of xenografts to examine these paracrine interactions in drug discovery applications, we will model the tumour microenvironment in vitro, thereby aligning with the objectives of the NC3Rs. To test the utility of this approach we will model a colon liver metastasis niche using early-passage patient-derived epithelial cells and matched CAFs in a biomatrix that has been shown to support multiple cell types in co-culture, using a functional HGF/C-met pathway as a measure of success. We will apply the refinement of real-time fluorescent/bioluminescent reporters to assess expansion of the different cell-types within the culture and monitor environmental signals such as hypoxia, EMT and apoptotic potential concurrently, without the need to harvest the cells. These will be validated against fresh human colorectal liver metastasis samples. Furthermore replacement of CAFs with commercially-available human mesenchymal stem cells (MSCs) will also be assessed as use of these should enable broader application of this in vitro system to cells derived from different tumour types. The final assessment of drug response will be performed in a 96-well format to examine the utility of this in vitro approach to a higher through-put format maximising its uptake by the pharmaceutical industry in drug discovery applications. This refined in vitro system therefore has the potential to replace the need for animals in investigating the biology of the tumour micro-environment for both new target identification and lead optimisation.

Impacts

Publications

  1. Musah-Eroje A and Watson S (2019). A novel 3D in vitro model of glioblastoma reveals resistance to temozolomide which was potentiated by hypoxia. JNeurooncol. 142:231-240. doi: 10.1007/s11060-019-03107-0
  2. Musah-Eroje A and Watson S (2019). Adaptive Changes of Glioblastoma Cells Following Exposure to Hypoxic (1% Oxygen) Tumour Microenvironment. Int. J. Mol. Sci. 20(9):2091 doi: 10.3390/ijms20092091
  3. Ahmed EM et al. (2018). A HIF-independent, CD133-mediated mechanism of cisplatin resistance in glioblastoma cells. Cell Oncol. 41:319. doi: 10.1007/s13402-018-0374-8
  4. Ivanov DP et al. (2018). In Vitro Tissue Microarrays for Quick and Efficient Spheroid Characterization. SLAS Discovery 23(2):211-217. doi: 10.1177/2472555217740576
  5. Ivanov DP et al. (2017). High-Throughput Spheroid Screens Using Volume, Resazurin Reduction, and Acid Phosphatase Activity. In: Gilbert D., Friedrich O. (eds) Cell Viability Assays. Methods in Molecular Biology, vol 1601. Humana Press, New York, NY
  6. Ivanov DP et al. (2017). Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D. Scientific Reports 7:41160. doi: 10.1038/srep41160
  7. Saunders JH et al. (2017). Individual patient oesophageal cancer 3D models for tailored treatment. Oncotarget 8(15):24224-242. doi: 10.18632/oncotarget.12500
  8. Ivanov DP et al. (2016). In vitro models of medulloblastoma: Choosing the right tool for the job. Journal of Biotechnology 236:10-25. doi: 10.1016/j.jbiotec.2016.07.028
  9. Ivanov DP et al. (2016). Separating chemotherapy-related developmental neurotoxicity from cytotoxicity in monolayer and neurosphere cultures of human fetal brain cells. Toxicology in vitro 37:88-96. doi: 10.1016/j.tiv.2016.09.007
  10. Onion D et al. (2016). 3-Dimensional Patient-Derived Lung Cancer Assays Reveal Resistance to Standards-of-Care Promoted by Stromal Cells but Sensitivity to Histone Deacetylase Inhibitors. Molecular Cancer Therapeutics 15(4):753-63. doi: 10.1158/1535-7163.MCT-15-0598